C-ring substituted 6alpha-halo steroids and process for producing them



like), nitrogen heterocyclics being preferred.

United States Patent Ofifice 3,079,384 Patented Feb. 26, 1953 3,079,384 (l-RING IJBSTITUTED tie-HALS STEROIDS AND PROQESS FUR PRQDUCHNG THEM Patrick A. Diassi, Westfieid, and Pacifier: A. Principe, New

Brunswick, N..'., assignors to Olin Mathieson Chemieai Corporation, New York, N.Y., a corporation of Virginia No Drawing. Filed Oct. 10, 19%, er. No. 61,388 19 Claims. (Cl. 260-23355) This invention relates to the preparation of new compounds which are primarily of advantage in the synthesis of C-ring substituted 6a-halo steroids and to methods of producing such compounds.

In general this invention involves the conversion of compounds of the formula (1) onion by a series of steps described in detail below by way of new intermediates ultimately to compounds of the formula The symbols in Formulas I and II have the following meanings: R and R" each represents hydrogen, lower alkyl (e.g. straight or branched chain aliphatic hydrocarbon groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, etc.), halogenated lower alkyl (e.g. trifluoromethyl or 2,2,2-trifluoroethyl), monocyclic cycloalkyl, (e.g. cyclopentyl, cyclohexyl etc.), monocyclic aryl (e.g. phenyl or mor ptoly1) and monocyclic aralkyl (e.g. phenyl lower alkyl such as benzyl or phenethyl); R represents hydrogen or an acyl radical of a hydrocarbon carboxylic acid, preferably of less than carbon atoms (e.g. lower alkanoic acids such as acetic, propionic,

'enanthic acids and the like), a monocyclic aryl carboxylic acid (e.g. benzoic acid), a monocyclic aralkanoic acid (e.g. a phenyl lower alkanoic acid such as phenylacetic 'acid, etc.), a cycloalkanoic or cycloalkenoic acid containing 3 to 6 carbon atoms in the ring, a monocyclic aralkenoic acid (e.g. a phenyl lower alkenoic acid such as cinnamic acid), or a lower alkenoic acid; X and X each represents a halogen, especially chlorine and fluorine,

which may be the same or different in an individual compound; and P and Q are independently selected from the group which consists of the radicals represented by R and R" (i.e. hydrogen, lower alkyl, halogenated lower alkyl, monocyclic cycloalkyl, monocyclic aryl, or monocyclic aralkyl), as well as monocyclic heterocyclic e.g. piperidine, piperazino, 4-methylpiperazino and pyrrolidino) or monocyclic heterocyclic lower alkyl (e.g. piperidinomethyl, pyrrolidinomethyl, furfuryl, thenyl and tcliile In ad tion, R' and R or P and Q may be joined together in an alkylene group of 3 to 5 carbon atoms or a monoazaalkylene group of 3 to 4 carbon atoms forming a cyclic hydrocarbon or nitrogen heterocyclic group together with the carbon to which they are attached.

In Formula II, the 1- and 2-positions may be saturated or they may be joined by a double bond. In other words the end products to which the intermediate of this invention leads are compounds of Formula II with positions 1 and 2 saturated as well as their 1,2-dehydro analogs.

In the first step of the process, which constitutes a principal feature of this invention, a compound of Formula I is subjected to the action of a microorganism of the genus Colletotrichum, 6g. C. phomoia'es, C. pisi, C. linicolae, etc., or the genus Tricothecium, e.g. T. roseum, T. flagrans, T. domestz'cum, etc. Colletotrichum phamoides and Tricothecium roseum are preferred. This results in the selective introduction of an lla-hydroxy group to yield a corresponding compound of the formula (m) GHZOH R, R" and X have the same significance in Formula III as they have in Formula I.

The hydroxylation can best be effected either by including the steroid of Formula I in an aerobic culture of the microorganism, or by bringing together, in an aqueous medium, the steroid, air and the microorganism. In general, the conditions for culturing the microor ganisms for the purposes of this invention are (except for the inclusion of the steroid to be converted) the same as those for culturing various other molds for the production of antibiotics and/or riboflavin.

The microorganism is aerobically grown in contact with (in or on) a suitable fermentation medium. A suitable medium essentially comprises a nitrogenous substance and a source of carbon and energy. The latter may be a carbohydrate (such as sucrose, molasses, glucose, maltose, starch or dextrin), a fatty acid, a fat and/or the steroid itself. Preferably, however, the medium includes an assimilable source of carbon and energy in addition to the steroid. The source of nitrogenous factors may be natural (e.g., soybean meal, corn steep liquor, meat extract and/or distillers solubles) or synthetic (i.e. composed of simple, synthesizable organic or inorganic compounds such as ammonium salts, alkali nitrates, amino acids or urea). An adequate sterile air supply should be maintained during fermentation, for example, by the conventional methods of exposing a large surface of the medium to air or by utilizing submerged aerated culture. The steroid may be added to the culture during the incubation period, or included in the medium prior to sterilization or inoculation. The preferred (but not limiting) range of concentration of the steroid in the culture is about 0.01 to 0.10% by weight. The culture period may vary considerably, e.g, within the range of about 6 to 96 hours.

In the fermentation step described above, it is desirable to use a starting material containing only hydrogen or hydrocarbon substituents in the l6,l 7-acetal or ketal moiety, i.e. R and R" represent hydrogen or a hydrocarbon group. If, however, a product is desired containing basic groups such as those answering the description of the symbols P and Q such groups may be introduced into the hydroxylation products of Formula III.

It is frequently convenient to carry out the Ila-hydroxylation step described above with an acetonide of Formula I, that is a compound of that formula in which R and R" each represents methyl. Then any other group desired in the 16,17-positions may be introduced after the fermentation procedure has been completed.

The conversion of the acetonide or similar group may be effected by hydrolyzing the compound of Formula III, e.g. by treating successively with formic acid, then sodium carbonate to produce a compound of the formula The compound of Formula IV is then reacted with an aldehyde or ketone in perchloric acid which bears the desired substituent groups corresponding to the formula The meaning of the symbols P and Q has already been defined. In this manner a compound having the formula (VI) CHQOH is readily obtained. 7

The final products of vFormula Ill include an additional halogen atom and, optionally, an additional double bond not in the starting materials of Formula Ior the 110:- hydroxylated compounds of Formulas Ill orVVI. These may be introduced into the molecule in optional sequence and thus several alternate routes may be followed after a compound of Formula Ill or Formula VI has been obtained.

According to one alternative, 21 double bond is introduced into the 1,2-position of the compound of Formula VI. This may be accomplished by enzymatic dehydrogenation with Bacterium cyclooxydans in accordance with the procedure of Example 1 of US. Patent No. 2,822,318. Acylation with an agent capable of converting the 21- hydroxy group to a 2l-acyloxy group, e.g. acyl halides,

preferably acyl chlorides, or acid anhydrides having acyl radicals corresponding to R yields a product of the formula onion (V III) 0 R20 R ed m as described above, then selectively dehydrogenating, e.g. by treating the latter with 2,3-dichloro-5,6-dicyanobenzoquinone.

The next step in the process entails the conversion of the compound of Formula VII to a compound of the formula (1X) onion P, Q, R and X have the significance already defined. R represents an alkylsulfonyl or monocyclic arylsulfonyl group, e.g. a lower alkylsulfonyl group such as methauesulfonyl, ethanesulfonyl, and the like, phenylsulfonyl or a substituted phenylsulfonyl group, especially lower alkylbenzenesulfonyl such as p-toluenesulfonyl or halobenzenesulfonyl such as p-bromobenzenesultonyl. Methanesulfonyl and p-toluenesulfonyl groups are preferred. Suitable sulfonylating agents which may be used are R-halides such as RTf-bromides and especially R"'- chlorides such as methanesulfonyl chloride and p-toluenesulfonyl chloride.

Compounds of Formula II in which the 9,11-position is double bonded are then prepared from compounds of Formula IX by treating the latter with an alkali metal salt of a lJwer fatty acid such as sodium acetate in a substantially anhydrous lower fatty acid medium such as acetic acid, treating the corresponding M -derivative thus formed, with an agent capable of effecting the addition of hypobromous acid to the 9,11 double bond, e.g. a mixture of a strong acid, such as perchloric, toluene sulfonic or trichloroacetic acids, an N-bromoamide or irnide such as N-bromoacetamide or N-bromosuccinimide and dioxane. The resulting :,11fi-b1011'l0h3ldfi11 is dehydrobrominated by treatment with a base such as sodium acetate or potassium acetate to the corresponding 9[3,ll;3- oxide. The latter is finally treated with a hydrohalide such as hydrogen chloride or hydrogen fluoride in an organic solvent medium to give the compounds of For mula II. If desired, the group R in position 21 and the acetal or ketal group in positions l6, l7 may be removed from the compounds of Formula II by cleaving, for example with formic acid, then hydrolyzing with alkali, for example potassium bicarbonate.

According to another modification of this invention a compound of Formula Vi is acylated to obtain a compound of Formula VIII as described above. The compound of Formula VH1 is then treated with a sulfonyiat- The symbols in the above formula have the same significance as described previously. Dehydrogenation of compounds of Formula X by chemical or microbiological means such as those already described yields also compounds of Formula IX which may be further processed as indicated above.

The compounds of Formula II are obtained from those of Formula X by treating the latter with an alkali metal salt of a lower fatty acid in a substantially anhydrous lower fatty acid medium, reacting the N -derivative thus obtained with an agent which effects the addition of hypobromous acid to the 9,11-dou-ble bond, dehydrobrominating the resulting 9a,1l,6-bromohydrin with an aqueous inorganic base to obtain the corresponding 9,8,115-oxide and finally reacting the latter with a hydrohalide in an organic solvent.

It is also possible, according to a third modification of this invention to oxidize compounds of Formula VIII e.g. with chromic anhydride-sulfuric acid in acetonedioxane to obtain compounds of the formula X These in turn may be selectively dehydrogenated, e.g. with 2,3-dichloro-5,6-dicyanobenzoquinone in dioxane, to obtain compounds of the formula (XIII) 01120 R in Which R and R" have the significance already defined, are used for this purpose. Monoperphthalic acid treatment of the ketal or acetal gives the 5a,6ot-oxide. Fission with boron trifiuoride or boron trichloride in ether benzene yields the corresponding 5a-hydroxy-6B-fiuoro or 5ahydroxy-GB-chloro compound which is converted to 60- fiuoro or 6u-chloro-l6u-hydroxycortexolone 16,17-ketal (or acetal) 21-acetate by chromium trioxide oxidation followed by treatment with anhydrous hydrogen chloride in acetone. The latter can the deacylated by hydrolysis with dilute alkali such as potassium carbonate or potassium hydroxide in the absence of oxygen to obtain the desired starting material.

The compounds of Formula II exhibit anti-inflammatory activity similar to that of cortisone and hydrocortisome and can thus be used in human and veterinary medicine for the treatment of conditions such as rheumatoid arthritis or adrenocortical hormone insufliciency, for which purpose they are administered in the same manner and in lieu of cortisone and hydrocortisone, the dosage being adjusted to account for the degree of activity of the individual steroid. Selected intermediates exert similar activity.

. The following examples are illustrative of the invention. All temperatures are expressed in degrees centigrade.

EXAMPLE 1 60t-FlMOr0-1 6 u-H y droxy e pi h ydrocortisone 16,1 7-Acet0nide Surface growth from a culture of T ricothecium roseum maintained on Gould agar of the following composition:

Grns. Agar 20 Glucose 10 Yeast extract 2.5 Potassium phosphate 1 Distilled water to 1 liter. Sterilized for 20 minutes at 15 p.s.i.

is suspended in 5 m'ls. of 0.01% Duponol solution [Merck Index, 7th edition (1960), p. 472]. 1 m1. portions of this suspension are used to inoculate eight 250 ml. Erlenmeyer flasks containing mls. of corn steep medium of the following composition:

The flasks are then incubated at 25 C. on a rotary shaker (280 cycles/min; 2 inch radius) for 72 hours after which a 10% by volume transfer is made from these flasks to each of seven two liter Erlenmeyer flasks containing 500 m'ls. of corn steep medium of the hereinbefore described composition. At the same time, there is added. to each flask 2.5 mls. of a mg./ml. solution of 6aiiuoro-l6a-hydroxycortexolone 16,17-acetonide in N,N- dimethylformamide (DMF). The flasks each having a final steroid concentration of 300 gamma/ml. are then incubated on a rotary shaker (280 cycles/minute; 2 inch radius).

Whole broth samples are taken for analysis at 24 hour intervals after the addition of the steroid. Each 8 ml. broth sample is extracted with 1 ml. of methyl isobutyl ketone (MIBK) and the MIBK extracts, spotted at 0.1 ml. and 0.2 ml. intervals on Whatman No. 1 paper are chromatographed on a toluene-propylene glycol system for three hours. The chromatograms are examined under UV. for the presence of the steroid substrate.

Four days after steroid addition, the broth from each Analysis.-Calcd for C H O F (436.50): C, 66.04; H, 7.62; F, 4.35. Found: C, 66.17; H, 7.36; F, 3.71.

EXAMPLE 2 16,1 7 -A cetophenone Derivative of 5oz-Ch10rO-16oc- H droxyepihydrocorzisone Following the procedure of Example 1 identically except for the substitution as steroid substrate of 6m-chloro- 16oz hydroxycortexolone 16,17 acetophenone derivative there is obtained as product, the 16,17-acetophenone derivative of 6ot-chloro-16a-hydroxyepihydrocortisone [6achloro-16,17 (a-methyl)benzylidenedioxyepicorticosterone].

EXAMPLE 3 6a-Fluoro-1dwHydr-oxyepihydrocortisone 16,17-Acet0nide Surface growth from a culture of Colletotrichum phomoides maintained on Gould agar of the following composition:

Gms. Agar, 20 Glucose 10 Yeast extract 2.5 Potassiumv phosphate 1 Distilled water to 1 liter sterilized for 20 minutes at psi.

is suspended in 5 mls. of 0.01% Duponol solution. 1 ml. portions of this suspension are used to inoculate eight 250 ml. Erlenmeyer flasks containing 50 mls. of a nutrient medium of the following composition:

Gms. Starch Malted cereal extract syrup 10 Peptone 20 Cerelose 44 NaNOg 3 KH JPQ; 1 KCl 0.5 MgSOJH O 0.5 PeSO 0.01

Tap water to 1 liter sterilized 20 minutes at 15 p.s.i.

The flasks are then incubated at C. on a rotary shaker (280 cycles/min; 2 inch radius) for 72 hours after which a 10% by volume transfer is made from these flasks to each of six two-liter Erlenmeyer flasks containing 500 11115. of medium of. the following composition:

Gms.

NaNO 3 Glucose 40 KH2PO'4 1 Agar 1 Tap water to 1 liter Sterilized 20 rnins,-at 15 p.s.i.

At the same time, there is added to each flask 2.5 mls. of 60 mg. per ml. solution of 6u-fiuoro-16u-hydroxycortexolone 16,17-acetonide in N,N-dimethylformamide (DMF). The flasks, each having a final steroid concentration of 300 gamma/m1, are then incubated on a rotary shaker (280 cycles/min; 2 inch radius).

Whole broth samples are taken for analysis at 24 hour intervals after the addition of the steroid. Each 8 ml. broth sample is extracted with 1 ml. of methyl isobutylketo-ne (It 113K) and the MIBK extracts, spotted at 0.1 ml. and 0.2 ml. intervals on Whatman No. 1 paper, are chromatographed on a toluene-propylene glycol system for three hours. The chromatograms are developed and examined under U.V. for the presence of the steroid substrate.

Seven days after steroid addition the broth from each of the flasks is filtered directly through a Buchner-Seitz clarifying pad apparatus with suction. The combined filtrates are extracted there times with 1000 ml. portions of MIBK chloroform. The MIBK extracts are combined, washed twice with 1500 ml. of water and evaporated to dryness in vacuo. The residue (about 991 mgs.) is crytallized from acetone-hexane to give about 285 mgs. of 60afluoro 160: hydroxyepihydrocortisone 16,17 acetom'de identical with the product of Example 1.

EXAMPLE 4 160a,] 7a-Cyclolzexylidene Derivative of 6oz-FluOi'O-16ct- H ydroxyepihydrocortisone Following the procedure of Example 1 of US. Patent No. 2,822,318, mg. of u-fluoro-l6a-hydroxyepihydrocortisone 16,17-acetonide is converted to the product fia-fluoro-16a-hydroxyepiprednisolone 16,17-acetonide.

In the same manner, the product of Example 2 is converted to the 16,17'acetophenone derivative of 6a-chlorot-hYdIOXYEPlPI'BdHlSOlOHE.

EXAMPLE 6 6a-Fluoro-I6a-Hydroxyepihydrocortisone 16,17- Acetonide 21 -Acetate To 100 mg. of 6a-fluoro-16u-hydroxyepihydrocortisone 16,17-acetonide are added 1.1 ml. of a solution of 108.7 mg. of acetic anhydride in 5 ml. of pyridine and the resulting solution is stoppered and left at room temperature for 16 hours during which time crystals separated. The solution is diluted slowly with 2 ml. of ice water and the crystals are filtered off, washed with water and dried to give about 78.6 mg. of a-fiuoro-l6a-hydroxyepihydrocortisone 16,17-acetonide 2l-acetate of the following characteristics: MP. about 293295; [Q]D+S8O (chlf.);

)-.i'1.237 my (e=15,50()); Ami? 2.80, 5.78, 5.86, 6.00, 6.161..

Analysis.Calcd for C i-$ 1 0 (478.54); C, 65.25; H, 7.37; F, 3.98. Found: C, 6479;1-1, 7.36; F, 3.61.

EXAMPLE 7 dot-Fluoro-ldot-Hydroxyepipredniso[one 21 -Acetate Following the procedure of Example 6, 6oc-flUOIO-16cchydroxyprednisolone 16,17-acetonide is acetylated to obtain oa-fluoro-l6a-hydroxyepiprednisolone 16,17-acetonide ZI-acetate.

16,17-Acetom'de EXAMPLE 8 dm-Fluoro Z 6 0t H ydroxy epihydrocortisone 1 6,1 7 -A cemnide 2] -Pr0pi0na. e and 6 Ot-C h Zora-1 6 ot-H ydroxyepihydrocortiroae 21 -A cemte 16,1 7-Ace10phen0ne Derivarive Following the procedure of Example 6 identically except for the substitution of 137 mgs. of propionic anhydride for the acetic anhydride used therein, there is obtained the product 6a-fiuoro-16e-hydroxyepihydrocortisone 16,17- acetonide 21-propionate.

in the same manner as in Example 6, the product of Example 2 is converted to the 16,17-acetophenone derivative of 6a-chloro-16a-hydroxyepihydrocortisone 21-acetate.

EXAMPLE 9 6 oz-F luoro-J 6 a-H ydroxyepihydrocortisone 11 -Mesylate 16,1 7-A cetonide 21 -A cetate To a stirred suspension of 50 mg. of 6a-fluoro-16ahydroxyepihydrocortisone 16,17-acetonide 21-acetate in 1.0 ml. of chloroform and 1.0 ml. of pyridine, cooled in an ice bath, is added 1.0 m1. of a solution of 0.1 ml. of methanesulfonyl chloride in 5.0 ml. of chloroform and the mixture is stirred at for 16 hours. The mixture is diluted with ml. of chloroform and then 3 ml. of ice water is added slowly. The chloroform layer is separated, washed successivelywith 2 N HCl, 5% NaHCO and water and then evaporated to dryness. Crystallization of the residue from acetone-hexane gives 6a-fluoro-16a-hydroxyepihydrocortisone ll-mesylate 16,17-acetonide 21- acetate, M.P. 175-177.

EXAMPLE 6a-Flu0r0-1oa-Hydroxyepihydrocortisone-ll -T0sy late 16,1 7-A cetonide 21 -A cetate Following the procedure of Example 9 except for the substitution of 50 mg. of tosyl chloride for the methanesulfonyl chloride solution used therein there is obtained the product 6a fiuoro 16a hydroxyepihydrocortisone ll-tosylate 16,17-acetonide 21-acetate.

EXAMPLE 11 16,17-Acet0phen0ne Derivative 0f 6oz-Chl0r0-16oz- H ydroxyepihydrocortisone 1 1 Ju es late 2] -A ce tale EXAMPLE 12 6a-Fluor0-16a-Hydroxyepiprednisolone 1 6 ,1 7 -A cetonide 21-Acetate A solution of 100 mg. of 6e-fiuoro-16oc-hyclroxyepihy- 'drocortisone 16,17-acetonide 21-acetate and 52 mg. of

2,3-dichloro-5,6-dicyanobenzoquinone in 2 ml. dioxane is refluxed under nitrogen for 6 hours. After cooling, the precipitated 2,3 dichloro-5,6-dicyanohydroquinone is filtered off and washed with 2 ml. dioxane. The filtrate and washings are diluted with 4 ml. of chloroform and adsorbed onto 2 g. of neutral alumina. Elution with chloroform and evaporation of the solvent gives 6a-fluoro-16ahydroxyepiprednisolone 16,17-acetonide 21-acetate.

EXAMPLE 13 6 :x-F [acre-16 a-H ydroxyepi prednisolone 1 6,1 7-A cetonia'e 21 -Pr0pi0nate Following the procedure of Example 12 except for the use as starting material of 6a-fluoro-loa-hydroxyepihydrocortisone 16,17-acetonide 21-propionate, there is obtained as product the compound 6oi-fluoro-l6a-hydroxyepiprednisolone 16,17-acetonide 21-propionate.-

EXAMPLE 14 16,17-Acetophen0ne Derivative 0 6oc-Chl0l'0-1 6a- H ydroxyepiprednisolone 21 -Acetate Following the procedure of Example 12 except for the use as starting material of the 16,17-acetophenone derivative of fia-chloro-16a-hydroxyepihydrocortisone 21-acetate there is obtained as product the 16,17-aceto-phenone derivative of 6a-chloro-16a-hydroxyepiprednisolone 2l-acetate.

EXAMPLE 15 6a Fluoro 16oz Hydroxyepiprednisolone 11 Mesylate 16,17-Acet0nia'e 21 -Acetate, 6e-Flu0r0-16a-Hydroxyepiprednisolone 16,17-Acet0nide 1 1 -Mesylate 21 -Pr0picrane and 6a-Chl0r0-1 6a-H ydroxyepiprednisolone 11- Mesylate 21 -Acetate 16,] 7-Acet0phen0ne Derivative To a stirred suspension of 50 mg. of 6u-fluoro-l6a-hydroxyprednisolone 16,17-acetonide 21-acetate in a mixture of 1 m1. of chloroform and 1 ml. of pyridine, cooled in an ice bath, is added 1.0 ml. of a solution of 0.1 ml. of methanesulfonyl chloride in 5.0 ml. of chloroform, and the mixture is kept at 0 for 16 hours. The mixture is diluted with 5 ml. of chloroform and 3 ml. of ice water. The chloroform layer is separated, washed with 2 N HCl, 5% NaHCO- and water and evaporated to dryness in vacuo. Crystallization of the residue from acetone-hexane gives 6a-fluoro-16a-hydroxyepiprednisolone l1-mesylate 16,17 acetonide 21-acetate.

In an identical manner the products of Examples 13 and 14 are converted to dot-fluoro-16a-hydroxyepiprednisolone 16,17-acetonide ll-mesylate 21-propionate and the 16,17- acetophenone derivative of a-chloro-l6a-hydroxyepiprednisolone ll-mesylate ZI-acetate respectively.

EXAMPLE 16 6a-F luoro-Z 6 e-H y droxyepiprednisolone 16,1 7 -Acet0nide l 1 -Mesy late 21 -A cetate In the same manner as in Example 12, the compound of Example 9 is converted to m-fiuoro-l6vx-hydroxyepiprednisolone 16,17-acetonide l1--nesylate 21-acetate.

EXAMPLE 17 6 oz-F luoro-l 6 a-H ydroxyepiprednisolone 1 1 -T0sy late- 16,17-Acet0nide 21-Acetate Following the procedure of Example 12, the product of Example 10 is converted to 6a-fluoro-16a-hydroxyepiprednisolone 1l-tosylate-16,17-acetonide 21-acetate.

EXAMPLE 18 16,17-Acetophenone Derivative of 6oc-Chl0rO-I6a- Hydroxyepiprednisolone 1 J-Mesylate 21 -Acetete Following the procedure of Example 12, the product of Example 11 is converted to 16,17-acetophenone derivative of 6a-chloro-16a-hydroxyepiprednisolone ll-mesylate 21- acetate.

EXAMPLE 19 6 a-Fluoro-l 6 a-H ydroxyepihydrocortisone A solution of 128 mg. of 6a-fiuoro-16a-hydroxyepihydrocortisone 16,17-acetonide 2l-acetate in 3.5 ml. of anhydrous formic acid is warmed at 42 for 22 hours. The solution is then evaporated to dryness in vacuo. The residue is dissolved. in 12.8 ml. of oxygen-free methanol and treated with 2.6 ml. of a 10% solution of potassium bicarbonate in water which is also free of oxygen. The reaction is left under nitrogen at room temperature for 30 minutes and then neutralized with 2.6 ml. of 10% acetic acid. Addition of 5 ml. of water followed by careful evaporation'of the methanol in vacuo gives crystalline 6o:- fluoro-l 6a-hydroxyepihydrocortisone.

EXAMPLE 2o Furfural Derivative of 6u-Flll0l0-16u- Hydroxyepihydrocorlisone neutralized with dilute sodium bicarbonate, water and chloroform are then addedahd the chloroform layer is 11 separated and concentrated in high vacuum. Crystallization of the residue gives the furfural derivative of 6c:- fluoro-16a-hydroxyepihydrocortisone.

EXAMPLE 21 da-Fluoro-I oa-Hydroxycortisone 1 6 ,1 7-A cetonide 21 -Acetate To a solution of 100 mg. of 6a-fiuoro-16a-hydroXyepihydrocortisone 16,17-acetonide 21-acetate in 3.0 m1. of reagent grade acetone, 1.0 ml. of an aqueous solution containing 20 mg. chromic'anhydride and 32 mg. sulfuric acid is added dropwise with stirring. After 10 minutes a few drops of methanol are added to decompose the excess oxidizing agent. The solution is decanted from the precipitated chrornic sulfate then diluted with water and eX- tracted with chloroform. The chloroform solution is washed with water and evaporated to dryness in vacuo. Crystallization of the residue from acetone-hexane gives 93.1 mg. of 6ct-fltl0IQ-16ot-hYdI'OXYCOIfiSOH8 16,17-acetonide Zl-acetate, M.P. 262-264",

k 234 mp. (e=l3,300)

Analysis.-Calcd for C H O F (476.32): C, 65.55; H, 6.99; F, 4.40. Found: C, 65.79; H, 6.86; F, 4.30.

EXAMPLE 22 6a-Flu0ro-16a-Hydroxyprednisone 16,1 7'Acet0nide 21-Aceta1e Following the procedure of Example 5, the product of Example 21 is converted to 6ot-flu0ro-16a-hydroxyprednisone 16,17-acetonide 21-acetate.

The invention may be otherwise variously embodied within the scope of the appended claims.

What is claimed is:

1. A compound of the formula CHaOR cmon O lower alhyl halogen the halogens being selected from the group consisting of chlorine and fluorine.

3. 6a-fiuor0-16a-hydroxyepihydrccortisone 16,17-acetonide.

16,17-ace- GHzOR wherein R represents a member of the group consisting of hydrogen and the acyl radical of a hydrocarbon car-boxylic acid or less than 10 carbon atoms, X represents halogen and P and Q each represents a member of the group consisting of hydrogen, lower alkyl, halo-lower alkyl, phenyl, phenyl lower alkyl, cycloalkyl, monocyclic heterocyclic, monocyclic heterocyclic lower alkyl, and, together with the carbon atom to which they are attached, a member of the group consisting of cycloalkyl and monocyclic heterocyclic, the halogens being selected from the group consisting of chlorine and fluorine.

7. A compound of the formula CHzO-Acyl 0 lower alkyl HO C 0 lower alkyl l halogen Acyl being the acyl radical of a hydrocarbon carboxylic acid of less than 10 carbon atoms and the halogens being selected from the group consisting of chlorine and fluorine.

8. 6a-fluoro-lwhydroxyepiprednisolone 16,17-acetonide.

9. 16,17-acetophenone derivative of 6ot-chloro-16u-hydroxyepiprednisolone.

10. 60c fiuoro 7 16oz hydroxyepiprednisolone 16,17- acetonide Zl-acetate.

11. A compound of the formula CHgOR wherein R represents a member of the group consisting of hydrogen and the acyl radical of a hydrocarbon carboxylic acid of less than 10 carbon atoms, X represents halogen and P and Q each represents a member of the 8,079,38 &

group consisting of hydrogen, lower alkyl, halo-lower alkyl, phenyl, phenyl lower alkyl, cycloalkyl, monocyclic heterocyclic, monocyclic heterocyclic lower alkyl, and, together with the carbon atom to which they are attached, a member of the group consisting of cycloalkyl and monocyclic heterocyclic, and R represents a member of the group consisting of alkylsulfonyl and monocyclic arylsulfonyl, the halogens being selected from the group consisting of chlorine and fluorine.

l2. 6a-fluoro-l6a-hydroxyepiprednisolone ll-mesylate 16,17-acetonide 2 l-acetate.

13. A compound of the formula CHHOR ""O P R/IIO I O: i

wherein R represents a member of the group consisting of hydrogen and the acyl radical of a hydrocarbon carboxylic acid of less than carbon atoms, X represents halogen and P and Q each represents a member of the group consisting of hydrogen, lower alkyl, halo-lower alkyl, phenyl, phenyl lower alkyl, cycloalkyl, monocyclic heterocyclic, monocyclic heterocyclic lower alkyl, and, together with the carbon atom to which they are attached, a member of the group consisting of cycloalkyl and monocyclic heterocyclic, and R' represents a member of the group consisting of alkylsulfonyl and monocyclic arylsulfonyl, the halogens being selected from the group consisting of chlorine and fluorine.

14. 6a-fiuoro-l6a-hydroxyepihydrocortisone ll-mesylate 16,17-acetonide Ill-acetate.

l5. 6a-fluoro-l6a-hydroxyepihydrocortisone ll-tosylate 16,17-acetonide Ill-acetate.

16. A process for the production of llu-hydroxy steroids of the formula CHaOR wherein R represents a member of the group consisting of hydrogen and the acyl radical of a hydrocarbon carboX- ylic acid of less than 10 carbon atoms, R and R" each represents a member of the group consisting of hydrogen, lower alkyl, halo-lower alkyl, phenyl, phenyl lower alkyl 14 a and cycloalkyl and X represents halogen, which comprises aerobically incubating a compound of the formula CHzOR 0 lower alkyl halogen which comprises aerobically incubating a compound of the formula CHROH I halogen in a nutrient medium comprising an assimilable source of nitrogen, carbon and energy in the presence of Colletotrichum phomoides.

19. A process for the production of 6oz-flu01O-16oc-11Y- droxyepihydrocortisone 16,17-acetonide which comprises aerobically incubating Ga-fluoro-l6ot-hydroxycortexolone a 16,17-acetonide in a nutrient medium comprising an assimilable source of nitrogen, carbon and energy in the presence of the microorganism Colletotrichum phomoides.

References Cited in the file of this patent UNITED STATES PATENTS Murray Feb. 16, 1960 OTHER REFERENCES Syntax, Belgium Patent No. 579,616 (reported in Derwent Belgian Report No. 60A, efiective date December 1959). 

1. A CPOMPOUND OF THE FORMULA 